Starfish Saponins, Part 28. Steroidal Glycosides from Pacific Starfishes

Nov 1, 1986 - Starfish Saponins, Part 28. Steroidal Glycosides from Pacific Starfishes of the Genus Nardoa. R. Riccio, O. Squillace Greco, L. Minale, ...
0 downloads 0 Views 217KB Size
Nov-Dec 19861

Brief Reports

1141

ACKNOWJ-XDGMENTS This work was supported by DIEXAT (Universidad de Antofagasta), Grant B-003, and FONDECYT, Grant 1053-84. The authors thank Professor C. Marticorena, University of Concepcion, for the identification of the plant material, Professor K. Naya, Kwansei Gakuin University, for kindly supplying a specimen of 6~-hydroxyeremophilanolideand 6 8 , 8~dihydroxyeremophilanolide,and Professor K. Yamada, Nagoya University for his generous gift ofsenkirkine. W e are indebted to Professor W.A. Ayer, Alberta University, for hrms measurements and to Professor F. Bohlmann, Technical University of Berlin, for running 400 MHz ‘H-nmr spectra. LITERATURE CITED 1. D. Yarnada, H . Tatematsu, Y.Kyotani, Y. Hirata, M. Haga, andT. Hirono, Phytochemistry, 17, 1667 (1978). 2. L. Novoty, M. Krojidlo, Z. Samek, J. Kohoutova, and F. Sorm, Collect. Czech. Chem. Commun., 38, 739 (1973). 3. K. Naya, R. Kanazawa, andM. Sawada, Bull. Chon. Soc.Jpn., 48,3220(1975). 4. A.R. Pinder, Progr. Chem. Org. Nat. Prod., 34, 8 1 (1977).

Reseived 25 Noyember 1985

STARFISH SAPONINS, PART 28. STEROIDAL GLYCOSIDES FROM PACIFIC STARFISHES OF THE GENUS NARDOA2 R. RICCIO,0. SQUILLACE GRECO,L. MINALE,+

Dipartimento di Chimica delle Sostanze Naturali, Universita, Via L. Rodind 22, 80138 Naples, ltaly D. DUHET,D. LAURENT,

Centre ORSTOM, B.P. AT, Noumba, New Caledonia

J. PUSSET,G. CHAUVIERE,and M. PUSSET Lahatoire des Plants MLdicinules, CNRS., B. P. 643 Noumk, New Caledonia Continuing our work on biologically active compounds from starfish (I), we have been working on glycosides of the Pacific starfishes Naraba nwaecaledonia Perrier and Narabagmphia Gray and have isolated several known steroidal glycosides previously found in Halityle regularis (2). From N . gompbia we have also isolated the sulfated glycosides, marthasteroside A, (3), thornasteroside A (4), and halityloside I (2), and two polyhydroxysteroids (25StSa-cholestane-38,6a,8, 158, 168,26-hexol (2), and (2&)-5a-cholestane-3f3,6a,8,158,24-pentol(5), all known compounds described from starfish. Table 1summarizes the results. EXPERIMENTAL ANIMAL COLLECTION AND EXTRACTION.-N. nwaecaledonia was collected in October 1983, and N . gomphia in March 1984, off NoumQ, New Caledonia, and identified by Mr. Michel Jangow of the Univenit6 Libre de Bruxelles. Samples of each species are deposited in the Centre ORSTOM, NoumQ. The starfish were chopped and soaked with H 2 0 for 4 h. The aqueous extracts were centrifuged and passed through a column of Amberlite XAD-2 (500 g for each column), which was washed with H,O (one bed volume) and then with MeOH. The MeOH eluates were taken to dryness to give 2.9 g of glassy material from N.nwaecaledonia and 4.7 g from N.gomophia.

‘For Part 27, see C. Pizza, L. Minale, D. Laurent, and J.L. Menou, Gazz. Chim. Itat., 115,(1985). ’This contribution is part of the Progetto Finalizzato “Chimica Fine e Secondaria” CNR, Rome, Italy.

1142

Wol. 49, No. 6

Journal of Natural Products

ISOLATION OF GLYCOSIDES AND POLAR STEROIDS.-For N . nwu&zf&nhz, the MeOH eluate was chrornatoghraphed on Sephadex LH-60 ( 4 x 8 0 cm, 100 g) with MeOH-H,O (2:l) (flow rate 13 mVh). Fractions (9 ml) were collected; fractions 50-60 contained 350 mg of a complex mixture of sulfated material (not further investigated); fractions 6 1- 130 contained steroidal glycosides (960 mg) and other compounds, and fractions 13 1-160 contained a mixture of tryptophan and tryptamine (85 mg), identified by comparison (tlc and 'H nmr) with authentic compounds. The glycoside fraction was further chromatographed on Sephadex LH-20 (2 X 60 cm; eluent, MeOH; flow rate 32 mYh; fractions of 8 ml were collected) to give 256 mg ofpartially purified glycosides (fractions 53-65), which were submitted to droplet countercurrent chromatography [dccc; CHCI3-MeOH-H,O (7: 13:8) in the ascending mode at a flow of 25 mYh; fractions of 5 ml were collected]. Fractions 55-75 contained halityloside A; fractions 80-103 contained halityloside B and fractions 122-150 contained halityloside D. Each of the above fractions was further purified by hplc on a p-bondapak C- 18 column (7.8 mm X 30 cm) using 25% H,O in MeOH. Quantities isolated are shown in Table 1.

TABLE1. Compounds Isolated from Nardoa Species I

I

I

Nardoa gornopbiab Reference (mg)

Compound

Halityloside A . . . . . . . . . . . . . -3.0" -5.0" Halityloside B . . . . . . . . . . . . . Halityloside D . . . . . . . . . . . . . - 13.0" Halityloside E . . . . . . . . . . . . . -20.4" Halityloside H . . . . . . . . . . . . . -4.7" Marthasteroside A, . . . . . . . . . . . +3.3" Thornasteroside A . . . . . . . . . . . . +5.3" Halityloside I . . . . . . . . . . . . . . (24S)-5a-cholestane-3p, 6a,8,15 p,24pentol . . . . . . . . . . . . . . . . -t 12.5' (25S>5a-cholestane-3p ,6a,8,15p, 168,262 0" hexol . . . . . . . . . . . . . . . .

813 797 783 767 827 1435 1289 883

18 13 14

15 15 5 30 5

19 12 7 15

(5)

5

"From 6 kg fresh material. bFrom 5.8 kg fresh material. For N. gornopbza, a similar isolation procedure was used. Chromatography of the MeOH eluate on Sephadex LH-60 gave two main fractions. The first fractions (25-45) contained the crude sulfated "asterosaponins" (650 mg), which were submitted to dccc [n-BuOH-Me,CO-H,O (3: 1:5) in the descending mode at a flow of 18 mVh; fractions of 7 ml were collected] to give, in fractions 92- 119,50 mg of a mixture of marthasteroside A, and thornasteroside A, which were finally separated by hplc on a p-bondapak C- 18 using 55% H,O in MeOH. The successive fractions (46-87) (1.9 g) contained a mixture of steroidal glycosides, polyhydroxysteroids, halityloside I, and other materials. These fractions were submitted to dccc [CHC13-MeOH-H20(7: 13:8) in the descending mode; flow rate 18 mYh; fractions of 3 ml were collected] and afforded in the more polar fractions (2 11-258) halityloside I, which was further purified by hplc on a p-bondapak C-18 column with 45% H,O in MeOH. The less polar fractions (23-162,346 mg) were further submitted to dccc with the same solvent system as above but in the ascending mode (fractions of 5 ml were collected) and afforded fractions, which were then submitted to hplc on a pbondapak C- 18 column with 25% H 2 0 in MeOH to give pure compounds. IDENTIFICATIONS.-Identification of each compound was based on hplc, optical rotations, 'H nmr and fabms and comparison with authentic samples. In the caseofhalitylosides A, B, D, and E, the 13C-nmr spectra were also determined. Full details of the identifications are available on request to the senior author. ACKNOWLEDGMENTS We thank the divers G. Bargibant, J.L.Menou, and P. Tirard of the Centre ORSTOM de Nourn& for collection of the animals ( N . gmpbia). We also thank Miss D. M a s c a g ~for her help in the isolation work.

Nov-Dec 19861

Brief Reports

1143

LITERATURE CITED 1. 2. 3.

4. 5.

L. Minale, R. Riccio, C. Pizza, and F. Zollo, “Proceedings of the Naito Foundation International Symposium on Natural Products and Biological Activities,” Tokyo, November 5-7, 1984. M. Iorizzi, L. Minale, R. Riccio, M. Debray, and J.L. Menou,]. Nut. Prod., 4 9 , 6 7 (1986). I. Bruno, L. Minale, C. Pizza, F. Zollo, R. Riccio, and F.A. Mellon, J . Chem. Sor. Perkin Trans I, 1875 (1984). I. KitagawaandM. Kobayashi, Chem. Phann. Bull., 26, 1864 (1978). R. Riccio, M.V.D’Auria, M. Ioriui, L. Minale, D. Laurent, and D. M e t , Gazz. Chim. It., 115, 405 (1985).

Rereiued I 3 December 1985

CHEMICAL COMPOSITION OF SANTOLZNA CHAMAECYPARZSSUS SSP. SQUARROSA ESSENTIAL OIL A. VILLAR,+R.M. GINER,and J.L. Rios Departamento de Farmacognosia y Farmacodinamia, Facultad de Farmacia, Uniuersidhd de Valencia, Avda. Blasco Zbairez 13, 46010 Valencia. Spain Santolina chamaecyparissusL. ssp. squarrosa DC.(Asteraceae) grows in eastern Spain. This plant is an aromatic dwarf shrub commonly known as “Manzanilla de Mahon” or “Abrotano hembra.” The flower of this plant is used in folk medicine because of its antispasmodic, digestive, antiinflamatory, antiseptic, and antimicrobial properties (1). Several studies concerning the composition of the essential oil from the species have been reported, but none have been about the essential oil fromsquarrosa ssp., which is endemic in the western Mediterranean area. This paper reports on the chemical composition ofS. chamaqparissus ssp. squarrosaessential oil. EXPERIMENTAL

PLANTMATERIAL-Fresh leaves and flowers of S . chamaecypatirsus ssp. squamsa were collected in Ayora-Enguera (Valencia) in June 1983. A voucher specimen has been deposited at the Department of Botany, Faculty of Pharmacy, University of Valencia. EXTRACTION OF ESSENTIAL orL.-Fresh plant material was subjected to steam distillation for 2.5 h using a modified Clevenger apparatus which yielded a yellowish essential oil (0.4% v/w). LIQUIDSOLID CHROMATOGRAPHY (Isc).-The oil was fractionated using Isc on a column (3.5 x 75 cm) ofdeactivated silica gel Merck (70-230 mesh ASTM) by addition of 5% H,O followed by gradient elution with hexane, mixtures of hexane/CH,CI,, and CH,CI, in order to separate the hydrocarbons and the oxygenated components of the oil. GAS-LIQUIDCHROMATOGRAPHY @IC).-The oil and its fractions obtained by Isc were analyzed by glc. Glc was performed using a Hewlett-Packard 5830 A gas chromatograph, equipped with FID, coupled to a 18850 A H-P data integrator. Conditions were as follows: High performance capillary column 5% OV-17 (25 mX0.20 mm) programmed from 80-150’ (rate 4’/min); the carrier gas was nitrogen (split technique ratio 1: 100); injection 225’; FID 250’. GAS CHROMATOGRAPHY-MASS SPECTROMETRY @c/ms).--Gc/ms was performed using an H-P 5995 gas chromatograph mass spectrometer with a membrane separator coupled to an H-P 9825 B control system. Conditions were asfollows: High performancecapillary 5% OV-17 (25 mX0.20 mm) was used in the same conditions as reported above for glc analyses; the carrier gas was helium (split technique ratio ca. 1: 100); electron energy 70 eV; ion source temperature 150’. Glc analysis showed the presence of 50 components: 39 compounds were identified, of which 28 corresponded to the monoterpenic fraction and 11 to the sesquiterpenic fraction. The essential oil contained a higher proportion of oxygenated compounds than hydrocarbons. Camphor (25.19%), p-cymene, 1,8cineole, bornyl and isobornyl acetate, allo-aromadendrene, and a-muurolene are the main constituents, totalling 68.11% ofthe essential oil. Twosesquiterpene alcohols were detected (M+=222 and MC=220), but they have not been identified. Identified compounds and their peak area percentages are listed in Table 1 according to their order of elution.